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ISSR (inter-simple sequence repeat) primer and method for identifying Dendrobium officinale Kimura et Migo and Dendrobium huoshanense C. Z. Tang et S. J. Cheng

A technology of Dendrobium officinale and Dendrobium officinale, applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve problems such as slow growth cycle of Dendrobium officinale, counterfeiting of Dendrobium officinale, harsh growth conditions, etc., and achieve Easy to quickly analyze and detect, less operation steps, good effect of polymorphic characteristics

Inactive Publication Date: 2018-09-28
湖北省农业科学院中药材研究所
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Problems solved by technology

The growth cycle of Dendrobium officinale is slow, the growth conditions are harsh, and the market demand is large, so counterfeit prod

Method used

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  • ISSR (inter-simple sequence repeat) primer and method for identifying Dendrobium officinale Kimura et Migo and Dendrobium huoshanense C. Z. Tang et S. J. Cheng
  • ISSR (inter-simple sequence repeat) primer and method for identifying Dendrobium officinale Kimura et Migo and Dendrobium huoshanense C. Z. Tang et S. J. Cheng
  • ISSR (inter-simple sequence repeat) primer and method for identifying Dendrobium officinale Kimura et Migo and Dendrobium huoshanense C. Z. Tang et S. J. Cheng

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Example Embodiment

[0031] Example 1 Establishment and optimization of ISSR-PCR amplification system

[0032] 1. Extraction of DNA

[0033] Rinse the mortar with water and dry it, add 5ml of absolute ethanol and light it. After the mortar is allowed to cool, take the collected Dendrobium officinale and 500mg of Dendrobium huoshanense buds in the mortar, pour an appropriate amount of liquid nitrogen to It is advisable to immerse the sample in liquid nitrogen by 2 to 3 cm in height, grind it for 3 to 5 minutes, and then extract the Dendrobium genomic DNA using the kit method.

[0034] 2. Establishment of ISSR-PCR amplification system

[0035] 1) Uniform design test plan of ISSR-PCR reaction system

[0036] Adopt U 12 (4 3 ) Uniform design table, the factor level of PCR amplification system is shown in Table 2. Using UBC834 as a primer and Dendrobium officinale genomic DNA as a template, the 12 reaction systems in Table 3 were used to optimize ISSR-PCR screening. The reaction system was 20 μL, as shown in T...

Example Embodiment

[0076] Example 2

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Abstract

The invention discloses an ISSR (inter-simple sequence repeat) primer and a reaction system for identifying Dendrobium officinale Kimura et Migo and Dendrobium huoshanense C. Z. Tang et S. J. Cheng. The nucleotide sequence of the primer is as shown in SEQ ID NO.1-6, and DNA (deoxyribonucleic acid) of the Dendrobium officinale Kimura et Migo and the Dendrobium huoshanense C. Z. Tang et S. J. Chengis amplified by the aid of the primer and an optimized reaction system, so that identification can be completed. The primer and a method are simple, time and cost are saved, and the primer has good application value to identification, genetic relationship analysis and the like of the Dendrobium officinale Kimura et Migo and the Dendrobium huoshanense C. Z. Tang et S. J. Cheng.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and in particular relates to an ISSR primer and a reaction system for identifying dendrobium officinale and dendrobium huoshanense. Background technique [0002] Dendrobium medicinal materials, as precious traditional Chinese medicines, have a long history of medicinal use and a wide range of sources, and the phenomenon of adulteration is extremely prominent. Among them, Dendrobium Huoshanense and Dendrobium officinale are often confused with each other due to the similarity in shape and structure. Dendrobium officinale Kimura et Migo is the most rare and precious species of Dendrobium officinale. The medicinal parts are fresh or dried stems, which have the functions of nourishing the stomach and promoting body fluid, nourishing yin and clearing heat, moistening the lungs and relieving cough, improving eyesight and strengthening the body. Dendrobium huoshanense (Dendrobium huoshanenseC...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 段媛媛游景茂卢超郭杰郭晓亮周武先
Owner 湖北省农业科学院中药材研究所
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